Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer
Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer
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DOI：
10.1002/jcb.28370
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发表时间：
2019-06-01
影响因子：
4
通讯作者：
Abolhassani, Mohsen
中科院分区：
文献类型：
--
作者：
Hassani, Mahmoud;Taheri, Fatemeh Hajari;Abolhassani, Mohsen
表达嵌合抗原受体（汽车）的T细胞的连续转移被认为是一种新的抗癌疗法。迄今为止，在大多数情况下，鼠来源的单链可变片段（scFv）已用于汽车。然而，这种结构具有与小鼠抗原在人中的潜在免疫原性和scFv的相对大的尺寸有关的限制。我们首次使用骆驼纳米抗体（VHH）构建了针对前列腺特异性膜抗原（PSMA）的CAR T细胞。针对PSMA（NBP）的纳米抗体用于显示CAR T细胞针对前列腺癌细胞的可行性。转染T细胞，然后确认CAR T细胞的表面表达。然后，在与前列腺癌细胞共培养后评价VHH-CAR T细胞的功能。最后，通过测定在遇到PSMA后CD 107 a的细胞表面表达来近似NBPII-CAR在T细胞中的细胞毒性潜力。我们的数据显示VHH-CAR T细胞对PSMA（+）细胞（LNCaP）的特异性，不仅通过增加白细胞介素2（IL-2）细胞因子（约400 pg/mL），而且通过增加几乎38%的CD 69表达。此外，与PSMA阴性前列腺癌细胞（DU-145）相比，当与LNCaP共培养时，VHH-CAR T细胞增殖接近60%，这导致T细胞中的CD 107 a上调高达31%。这些结果清楚地显示了使用基于VHH的CAR T细胞进行靶向免疫治疗的可能性，其可以被开发为靶向几乎任何肿瘤相关抗原，用于实体瘤的过继性T细胞免疫治疗。
Adoptive transfer of T cells expressing chimeric antigen receptors (CARs) is considered to be a novel anticancer therapy. To date, in most cases, single-chain variable fragments (scFvs) of murine origin have been used in CARs. However, this structure has limitations relating to the potential immunogenicity of mouse antigens in humans and the relatively large size of scFvs. For the first time, we used camelid nanobody (VHH) to construct CAR T cells against prostate specific membrane antigen (PSMA). The nanobody against PSMA (NBP) was used to show the feasibility of CAR T cells against prostate cancer cells. T cells were transfected, and then the surface expression of the CAR T cells was confirmed. Then, the functions of VHH-CAR T cell were evaluated upon coculture with prostate cancer cells. At the end, the cytotoxicity potential of NBPII-CAR in T cells was approximated by determining the cell surface expression of CD107a after encountering PSMA. Our data show the specificity of VHH-CAR T cells against PSMA(+) cells (LNCaP), not only by increasing the interleukin 2 (IL-2) cytokine (about 400pg/mL), but also the expression of CD69 by almost 38%. In addition, VHH-CAR T cells were proliferated by nearly 60% when cocultured with LNCaP, as compared with PSMA negative prostate cancer cell (DU-145), which led to the upregulation of CD107a in T cells upto 31%. These results clearly show the possibility of using VHH-based CAR T cells for targeted immunotherapy, which may be developed to target virtually any tumor-associated antigen for adoptive T-cell immunotherapy of solid tumors.